Construction and expression of the recombinant plasmid containing BddhFVIII in HepG2 cells / 中国应用生理学杂志

ABSTRACT

<p><b>OBJECTIVE</b>To get stable cell line expressing B domain-deleted human FVIII (BDDhFVIII) by constructing the eukaryotic expression plasmid.</p><p><b>METHODS</b>Eukaryotic expression plasmid containing BDDhFVIII was constructed and transfected into HepG2 cells via electroporation. The expression and purification of the target protein was detected by Western blot.</p><p><b>RESULTS</b>Results of enzyme digestion and sequence analysis demonstrated that the gene of BDDhFVIII was correctly inserted into the eukaryotic expression vector pcDNA4/v5-his. Western blot confirmed the successful expression of BDDhFVIII at the protein levels in HepG2 cells.</p><p><b>CONCLUSION</b>The constructed eukaryotic expression vector was able to generate high level expression of human FVIII in HepG2 cells, thus could construct human blood coagulation FVIII stable cell line successfully.</p>